Molecular Biotechnology

, Volume 54, Issue 2, pp 451–460 | Cite as

Efficient Regeneration Potential is Closely Related to Auxin Exposure Time and Catalase Metabolism During the Somatic Embryogenesis of Immature Embryos in Triticum aestivum L

  • Maoyun She
  • Guixiang Yin
  • Jiarui Li
  • Xing Li
  • Lipu Du
  • Wujun Ma
  • Xingguo Ye
Research
First Online:

Abstract

Regeneration of cultured tissue is a prerequisite of Agrobacterium- and biolistic-mediated plant transformation. In this study, an efficient protocol for improving wheat (Triticum aestivum L.) immature embryo regeneration was developed. Based on the statistical analysis of embryogenic callus induction efficiency, green spot differentiation efficiency, and plant regeneration efficiency from five wheat accessions, improved culture conditions were found to be more effective for embryogenic callus production than the traditional conditions. Using semi-quantitative reverse transcription polymerase chain reaction, a candidate gene, designated as TaCAT1, which encodes a catalase was identified to have a significant correlation with high-regeneration trait of wheat immature embryos. Three amino acid substitutions were found in TaCAT1 protein between high- and low-regeneration wheat accessions. Hydrogen peroxide content in the cultured calli increased from day 5 to 15, and then decreased sharply on day 20, followed by a second peak on day 25 during regeneration stage. Furthermore, a 3,500-bp 5′ flanking region upstream of the first codon ATG of TaCAT1 was isolated using inverse polymerase chain reaction. In silico, analysis revealed that the TaCAT1 promoter contained two regulatory motifs associated with responses to auxin.

Keywords

Triticum aestivum Immature embryo culture Regeneration capacity TaCAT1 Hydrogen peroxide content 
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Notes

Acknowledgments

This research was financially supported by the National Natural Science Foundation of China (Grant No. 30971776) and the Ministry of Agriculture of China (Grant No. 2008ZX08010-004).

Supplementary material

12033_2012_9583_MOESM1_ESM.pdf (338 kb)
Additional supporting information may be found in the online version of this article: Figure S1, Figure S2, Figure S3, and Table S1, Table S2 (PDF 337 kb)

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Maoyun She
    • 1
  • Guixiang Yin
    • 1
  • Jiarui Li
    • 3
  • Xing Li
    • 1
  • Lipu Du
    • 1
  • Wujun Ma
    • 4
  • Xingguo Ye
    • 1
    • 2
  1. 1.Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, National Key Facility of Crop Gene Resources and Genetic Improvement, Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijingChina
  2. 2.National Key Facility of Crop Gene Resources and Genetic ImprovementChinese Academy of Agricultural SciencesBeijingChina
  3. 3.Department of Plant PathologyKansas State UniversityManhattanUSA
  4. 4.Department of Agriculture & Food, Centre for Comparative GenomicsMurdoch University & Western AustralianPerthAustralia

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